Anemoside B4 mitigates endoplasmic reticulum stress-induced apoptosis post cerebral ischemia/reperfusion injury in rats

Articles in Press

Document Type : Original Article

Authors

1 Department of Pathology, Chongqing Three Gorges Medical College, Wanzhou, China

2 Chongqing Three Gorges Medical College, Wanzhou, China

3 Medical college, Jiujiang University, Jiujiang, Jiangxi Province , China

10.22038/ijbms.2025.85441.18475

Abstract

Objective(s): Anemoside B4 (AB4) exhibits neuroprotective effects on cerebral ischemia/reperfusion injury (CIRI), and endoplasmic reticulum stress (ERS) plays a crucial role in the process of CIRI. Nevertheless, it remains unknown whether AB4 acts on CIRI via ERS. This study is designed to determine whether AB4 mitigates CIRI by suppressing ERS-induced neuronal apoptosis.
Materials and Methods: One hundred thirty-five male SD rats (260–290 g) were randomly assigned to five groups, with 20 rats in each group: ① Sham. ② Middle Cerebral Artery Occlusion (MCAO/R). ③AB4-L: Prior to MCAO, rats were subjected to continuous intraperitoneal injection of 1.25 mg/kg of AB4.④AB4-M: Rats were administered a continuous intraperitoneal injection of 2.5 mg/kg of AB4 prior to undergoing MCAO.⑤AB4-H: Rats were continuously intraperitoneally injected with 5 mg/kg of AB4 before MCAO. Additionally, another thirty-five rats were employed for the time point. 
Results: TTC staining results demonstrated that AB4 significantly reduced cerebral infarct volume. Histopathological analysis of brain tissues revealed that AB4 mitigated neuronal damage. In the MCAO model, GRP78 expression progressively increased with reperfusion time and peaked at 24 hr. AB4 treatment decreased mRNA levels of key ERS markers, including GRP78, ATF6, IRE1α, and PERK. Additionally, AB4 reduced protein levels of GRP78, p-PERK, PERK, ATF6, and p-IRE1α, further indicating its role in attenuating ERS. TUNEL results demonstrated that AB4 significantly reduced neuronal apoptosis.
Conclusion: AB4 may serve as a potential therapeutic agent for CIRI, potentially exerting neuroprotective effects through inhibiting ERS-mediated apoptosis.

Keywords

Main Subjects

References

1. Hilkens NA, Casolla B, Leung TW, de Leeuw FE. Stroke. Lancet 2024; 403: 2820-2836.
2. Qin C, Yang S, Chu YH, Zhang H, Pang XW, Chen L, et al. Signaling pathways involved in ischemic stroke: molecular mechanisms and therapeutic interventions. Signal Transduct Target Ther 2022; 7: 215-244.
3. Zhang M, Liu Q, Meng H, Duan H, Liu X, Wu J, et al. Ischemia-reperfusion injury: Molecular mechanisms and therapeutic targets. Signal Transduct Target Ther 2024; 9: 12-51.
4. Chen X, Shi C, He M, Xiong S, Xia X. Endoplasmic reticulum stress: Molecular mechanism and therapeutic targets. Signal Transduct Target Ther 2023; 8: 352-392.
5. Ji X, Chen Z, Lin W, Wu Q, Wu Y, Hong Y, et al. Esculin induces endoplasmic reticulum stress and drives apoptosis and ferroptosis in colorectal cancer via PERK regulating eIF2alpha/CHOP and Nrf2/HO-1 cascades. J Ethnopharmacol 2024; 328: 118139.
6. Kang K, Chen SH, Wang DP, Chen F. Inhibition of endoplasmic reticulum stress improves chronic ischemic hippocampal damage associated with suppression of IRE1alpha/TRAF2/ASK1/JNK-dependent apoptosis. Inflammation 2024; 47: 1479-1490.
7. Marciniak SJ, Chambers JE, Ron D. Pharmacological targeting of endoplasmic reticulum stress in disease. Nat Rev Drug Discov 2022; 21: 115-140.
8. Wang L, Liu Y, Zhang X, Ye Y, Xiong X, Zhang S, et al. Endoplasmic reticulum stress and the unfolded protein response in cerebral ischemia/reperfusion injury. Front Cell Neurosci 2022; 16: 864426.
9. Wu MH, Hsieh YH, Lin CL, Ying TH, Hsia SM, Hsieh SC, et al. Licochalcone A induces endoplasmic reticulum stress-mediated apoptosis of endometrial cancer cells via upregulation of GRP78 expression. Environ Toxicol 2024; 39: 2961-2969.
10. Meng CC, Zhang JY, Zhang LY, Wang YT, Li ZY, Zhao J. Effects of NLRP6 in cerebral ischemia/reperfusion (I/R) injury in rats. J Mol Neurosci 2019; 69: 411-418.
11. Merighi A, Lossi L. Endoplasmic reticulum stress signaling and neuronal cell death. Int J Mol Sci 2022; 23: 15186-15216.
12. Huang X, Gan H, Tan J, Wang T, Zhao J, Zhao Y. BRCC3 promotes activation of the NLRP6 inflammasome following cerebral ischemia/reperfusion (I/R) injury in rats. Neurosci Lett 2021; 756: 135954.
13. Yuan R, He J, Huang L, Du LJ, Gao H, Xu Q, et al. Anemoside B4 protects against acute lung injury by attenuating inflammation through blocking NLRP3 inflammasome activation and TLR4 dimerization. J Immunol Res 2020; 2020: 7502301-7502314.
14. Fei H, Huang X. Effect of anemoside B4 on ameliorating cerebral ischemic/reperfusion injury. Iran J Basic Med Sci 2025; 28: 49-55.
15. Fei HZ, Xiang P, Luo W, Tan XD, Gu C, Liu MZ, et al. CTRP1 attenuates cerebral ischemia/reperfusion injury via the PERK signaling pathway. Front Cell Dev Biol 2021; 9: 700854-700871.
16. Zhou F, Wang YK, Zhang CG, Wu BY. miR-19a/b-3p promotes inflammation during cerebral ischemia/reperfusion injury via SIRT1/FoxO3/SPHK1 pathway. J Neuroinflammation 2021; 18: 122-135.
17. Hu L, Wei X, Shen G, Huang X. Ellagic acid alleviates NLRP6/caspase-1/GSDMD-mediated inflammation and pyroptosis in rats post cerebral ischemia/reperfusion injury. Iran J Basic Med Sci 2025; 28: 105-112.
18. Huang X, Tan J, Ji Y, Luo J, Zhao Y, Zhao J. BRCC3 mediates inflammation and pyroptosis in cerebral ischemia/reperfusion injury by activating the NLRP6 inflammasome. CNS Neurosci Ther 2024; 30: e14697-14711.
19. Elwakiel A, Mathew A, Isermann B. The role of endoplasmic reticulum-mitochondria-associated membranes in diabetic kidney disease. Cardiovasc Res 2024; 119: 2875-2883.
20. Ernst R, Renne MF, Jain A, von der Malsburg A. Endoplasmic reticulum membrane homeostasis and the unfolded protein response. Cold Spring Harb Perspect Biol 2024; 16: a041400-41431.
21. Newton K, Strasser A, Kayagaki N, Dixit VM. Cell death. Cell 2024; 187: 235-256.
22. Ni D, Lei C, Liu M, Peng J, Yi G, Mo Z. Cell death in atherosclerosis. Cell Cycle 2024; 23: 495-518.
23. Lasorsa F, Rutigliano M, Milella M, d’Amati A, Crocetto F, Pandolfo SD, et al. Ischemia-reperfusion injury in kidney transplantation: Mechanisms and potential therapeutic targets. Int J Mol Sci 2024; 25: 4332-4345.
24. Xiang Q, Yi X, Zhu XH, Wei X, Jiang DS. Regulated cell death in myocardial ischemia-reperfusion injury. Trends Endocrinol Metab 2024; 35: 219-234.
25. Zheng J, Li Y, Zhang T, Fu Y, Long P, Gao X, et al. Endoplasmic reticulum stress and autophagy in cerebral ischemia/reperfusion injury: PERK as a potential target for intervention. Neural Regen Res 2025; 20: 1455-1466.
26. Chen Y, Chen Z, Cheng W, Cao Y, Xu W, Lai W, et al. The role of the GRP78/PERK/ATF4 pathway in the ability of gua lou gui zhi decoction to attenuate apoptosis by inhibiting endoplasmic reticulum stress after ischemia-reperfusion injury. Front Biosci (Landmark Ed) 2022; 27: 296.
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 27 May 2025

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